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Traps and Pitfalls—Unspecific Reactions in Metabolic Engineering of Sesquiterpenoid Pathways

机译:陷阱和陷阱—倍半萜途径代谢工程中的非特异性反应

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摘要

The characterization of plant enzymes by expression in prokaryotic and eukaryotic (yeast and plants) heterologous hosts has widely been used in recent decades to elucidate metabolic pathways in plant secondary metabolism. Yeast and plant systems provide the cellular environment of a eukaryotic cell and the subcellular compartmentalization necessary to facilitate enzyme function. The expression of candidate genes in these cell systems and the identification of the resulting products guide the way for the identification of enzymes with new functions. However, in many cases, the detected compounds are not the direct enzyme products but are caused by unspecific subsequent reactions. Even if the mechanisms for these unspecific reactions are in many cases widely reported, there is a lack of overview of potential reactions that may occur to provide a guideline for researchers working on the characterization of new enzymes. Here, an across-the-board summary of rearrangement reactions of sesquiterpenes in metabolic pathway engineering is presented. The different kinds of unspecific reactions as well as their chemical and cellular background are explained and strategies how to spot and how to avoid these unspecific reactions are given. Also, a systematic approach of classification of unspecific reactions is introduced. It is hoped that this mini-review will stimulate a discussion on how to systematically classify unspecific reactions in metabolic engineering and to expand this approach to other classes of plant secondary metabolites.
机译:通过在原核和真核(酵母和植物)异源宿主中表达来表征植物酶,在最近几十年中已广泛用于阐明植物次生代谢中的代谢途径。酵母和植物系统提供了真核细胞的细胞环境和促进酶功能所必需的亚细胞区室化。这些细胞系统中候选基因的表达以及所得产物的鉴定为鉴定具有新功能的酶提供了指导。但是,在许多情况下,检测到的化合物不是直接的酶产物,而是由随后的非特异性反应引起的。即使在许多情况下已广泛报道了这些非特异性反应的机制,但仍缺乏对可能发生的潜在反应的概述,从而无法为研究新酶表征的研究人员提供指导。在这里,全面概述了倍半萜在代谢途径工程中的重排反应。解释了不同种类的非特异性反应及其化学和细胞背景,并给出了如何发现和避免这些非特异性反应的策略。此外,介绍了非特异性反应分类的系统方法。希望这份小型综述会激发有关如何在代谢工程中对非特异性反应进行系统分类并将这种方法扩展到其他类别的植物次生代谢产物的讨论。

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